In late 2002, cases of life-threatening respiratory disease with no как передается коронавирус у кошек identifiable etiology were reported from Guangdong Province, China, followed by reports from Vietnam, Canada, and Hong Kong of severe febrile respiratory illness that spread to household members and health care workers. The syndrome was designated “severe acute respiratory syndrome” (SARS) in February 2003 by the Centers for Disease Control and Prevention ( MMWR, 52:241–48, 2003).
Past efforts to develop rapid diagnostics and vaccines for coronavirus infection как передается коронавирус у кошек in humans have been hampered by a lack of appropriate research models and the moderate course of disease in humans. Therefore, a need for rapid diagnostic tests and vaccines exists. SUMMARY OF THE DISCLOSURE A newly isolated human coronavirus has been identified as the causative agent of SARS, and is termed SARS-CoV.
The nucleic acid sequence of the SARS-CoV genome and the amino acid sequences of the SARS-CoV open reading frames are provided herein. This disclosure provides methods как передается коронавирус у кошек and compositions useful in detecting the presence of a SARS-CoV nucleic acid in a sample and/or diagnosing a SARS-CoV infection in a subject. Also provided are methods and compositions useful in detecting the presence of a SARS-CoV antigen or antibody in a sample and/or как передается коронавирус у кошек diagnosing a SARS-CoV infection in a subject. The foregoing and other features and advantages will become more apparent from the following detailed description of several embodiments, which proceeds with reference to the accompanying figures. 1A–B are photomicrographs illustrating typical early cytopathic effects seen with coronavirus isolates and serum from SARS patients. 1A is a photomicrograph of Vero E6 cells inoculated with an oropharyngeal specimen from a SARS как передается коронавирус у кошек patient (?40). 1B is a photomicrograph of infected Vero E6 cells как передается коронавирус у кошек как передается коронавирус у кошек reacting with the serum of a convalescent SARS patient in an indirect fluorescent antibody (IFA) assay (?400). 2A–B are electronmicrographs illustrating ultrastructural characteristics of the SARS-associated coronavirus (SARS-CoV).
2A is a thin-section electron-microscopical view of viral nucleocapsids aligned along the membrane of the rough как передается коронавирус у кошек endoplasmic reticulum (arrow) as particles bud into the cisternae.
Enveloped virions have surface projections (arrowhead) and an electron-lucent center. Directly under the viral envelope lies a characteristic ring formed by the helical nucleocapsid, often seen in cross-section. 2B is a negative stain (methylamine tungstate) electronmicrograph showing stain-penetrated coronavirus particle with the typical internal helical nucleocapsid-like structure and club-shaped surface projections surrounding the periphery of the particle. 3 как передается коронавирус у кошек is an estimated maximum parsimony tree illustrating putative phylogenetic relationships between SARS-CoV and other human and animal coronaviruses. Phylogenetic relationships are based on sequence alignment of 405 nucleotides of the coronavirus polymerase gene ORF1b (nucleic acid 15,173 to 15,578 of SEQ ID как передается коронавирус у кошек NO: 1). The three major coronavirus antigenic groups (I, II and как передается коронавирус у кошек III), represented by HcoV-229E, CCoV, FIPV, TGEV, PEDV, HcoV-OC43, BCoV, HEV, SDAV, MHV, TCoV, and IBV-Avian, are shown shaded. Bootstrap values (100 replicates) obtained from a 50% majority rule consensus tree are plotted at the main internal branches of the phylogram. Branch lengths are proportionate to nucleotide differences. 4 is a pictorial representation of neighbor joining trees illustrating putative phylogenetic relationships between SARS-CoV and other human and как передается коронавирус у кошек animal coronaviruses. Amino acid sequences of the indicated SARS-CoV proteins were как передается коронавирус у кошек как передается коронавирус у кошек compared with those from reference viruses representing each species in the three groups of coronaviruses for which complete genomic sequence information was available [group 1: HCoV-229E (AF304460); PEDV (AF353511); TGEV (AJ271965); group 2: BCoV (AF220295); MHV (AF201929); group 3: infectious bronchitis virus (M95169)].
Sequences for representative strains of other coronavirus species, for which partial sequence information was available, were included for some of the structural как передается коронавирус у кошек protein comparisons [group 1: CCoV (D13096); FCoV (AY204704); porcine respiratory coronavirus (Z24675); group 2: HCoV-OC43 (M76373, L14643, M93390); HEV (AY078417); rat coronavirus (AF207551)]. Sequence alignments and neighbor-joining trees were generated by using Clustalx 1.83 with the Gonnet protein comparison matrix. The resulting trees were adjusted for final output using treetool 2.0.1. 5A–C are photomicrographs illustrating diffuse alveolar damage in a patient with SARS ( FIGS.
5A–B ), and immunohistochemical staining of SARS-CoV-infected Vero E6 cells ( FIG. 5A is a photomicrograph of lung tissue from a SARS patient (?50).
Diffuse alveolar damage, abundant foamy macrophages and multinucleated syncytial cells are present; hematoxylin and eosin stain was used. 5B is a higher magnification photomicrograph of lung tissue from the same SARS patient (?250).
Syncytial cells show no conspicuous viral inclusions.
5C is a как передается коронавирус у кошек photomicrograph of immunohistochemically stained SARS-CoV-infected cells (?250). Membranous and cytoplasmic immunostaining как передается коронавирус у кошек of individual and syncytial Vero E6 cells was achieved using feline anti-FIPV-1 ascitic fluid.
Immunoalkaline phosphatase with naphthol-fast red substrate and hematoxylin counter stain was used. 6A–B are electronmicrographs illustrating ultrastructural characteristics of a как передается коронавирус у кошек coronavirus-infected cell in bronchoalveolar lavage (BAL) from a SARS patient. 6A как передается коронавирус у кошек is an electronmicrograph of a coronavirus-infected cell. Numerous intracellular and extracellular particles are present; virions are indicated by the arrowheads. 6B is a higher magnification electronmicrograph of the area seen at the arrow in FIG. 7A–C illustrate the organization of the SARS-CoV genome. 7A is a diagram of the overall organization of the 29,727-nt как передается коронавирус у кошек SARS-CoV genomic RNA. The 72-nt leader sequence is represented as a small rectangle at the left-most end.
ORFs1a and 1b, encoding the nonstructural polyproteins, and those ORFs encoding the S, E, M, and N structural proteins, are indicated. Vertical position of the boxes indicates the как передается коронавирус у кошек phase of the reading frame (phase 1 for proteins above the line, phase two for proteins on the line and phase three for proteins below the line). 7B is an expanded view of the structural protein encoding region and predicted mRNA transcripts. Known structural protein encoding regions (dark grey boxes) and regions and reading frames как передается коронавирус у кошек как передается коронавирус у кошек for potential products X1–X5 (light gray boxes) are indicated. Lengths and map locations of the 3?-coterminal mRNAs expressed by the SARS-CoV are indicated, as predicted by identification of conserved transcriptional regulatory sequences.
7C is a digitized image of a nylon membrane showing Northern как передается коронавирус у кошек blot analysis of SARS-CoV mRNAs. Poly(A)+ RNA from infected Vero как передается коронавирус у кошек как передается коронавирус у кошек E6 cells was separated on a formaldehyde-agarose gel, transferred to a nylon membrane, and hybridized with a digoxigenin-labeled riboprobe overlapping the 3?-untranslated region. Sizes of the SARS-CoV mRNAs were calculated by extrapolation как передается коронавирус у кошек from a log-linear fit of the molecular mass marker. Lane 1, SARS-CoV mRNA; lane 2, Vero E6 cell mRNA; lane 3, molecular как передается коронавирус у кошек mass marker, sizes in kB. SEQUENCE LISTING The nucleic and amino acid sequences listed in the accompanying sequence listing are shown using как передается коронавирус у кошек standard letter abbreviations for nucleotide bases, and three letter code for amino acids, as defined in 37 C.F.R. Only one strand of each nucleic acid sequence is shown, but the complementary strand is understood as included by any reference to the displayed strand.
